Use a concentration of 1 µg/ml. Detects a band of approximately 155 kDa (predicted molecular weight: 129 kDa). Human Thrombospondin-1 is predicted to migrate in WB to ~155kDa as it contains a number of potential glycosylation sites; Swiss Prot predicts a protein of 129kDa not taking modifications into account.

ターゲット情報

関連性Thrombospondin is a regulator of many biological processes including cell growth, adhesion, migration, platelet aggregation, and fibrin deposition and lysis. It interacts with a number of plasma proteins such as fibrinogen and plasminogen and co-polymerizes with fibrin in clot formation. Thrombospondin also has multiple binding sites that interact with molecules such as fibronectin, collagens, laminin and heparan sulphate proteoglycans as well as binding growth factors such as TGF-beta1. Thrombospondin exerts an anti-adhesive effect which leads to cell rounding and detachment.

Anti-Thrombospondin 1 antibody 画像

IHC image of Thrombospondin staining in human breast adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab85762, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

Exposure time : 5 minutesHuman Thrombospondin-1 is predicted to migrate in WB to ~155kDa as it contains a number of potential glycosylation sites; Swiss Prot predicts a protein of 129kDa not taking modifications into account.

ICC/IF image of ab85762 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab85762, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) HepG2 cells at 5µg/ml, and in 100% Methanol fixed (5 min) HeLa, and HepG2 cells at 5µg/ml.

ab85762 used in Immunoprecipitation.Whole cell lysate prepared from human endothelial cells loaded at 600µg total protein.Immunoprecipitation step performed using Protein A.For IP: ab85762 used at 3.33µg/mg lysate.For WB: ab85762 used at a1/1000 dilution.